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Developing a DNA Extraction Method for Mineral Abundant Limestone Cave Samples
No full textCaves offer a unique environment to study microbial life that survives under extreme nutrient limitation. To this extent, 16s metabarcoding serves as a sort of “DNA fingerprint” that allows us to identify and study the microbial composition of these environments, but there are hurdles that must be overcome to obtain enough DNA for effective and reliable 16s sequencing. It is important to maximize the amount of DNA that can be extracted from the Wind Cave samples because it is an extremely low biomass environment and sufficient DNA for sequencing can be hard to get. Wind Cave National Park serves as a relatively accessible cave microbiome to study. It is a limestone karst cave, meaning it is largely made up of the mineral calcite, which can inhibit DNA extraction because of positive charges in the mineral structure trapping negatively charged DNA on its surface. Our lab has previously worked on developing a DNA extraction protocol suited for high DNA yield from environmental samples, including many experiments with Wind Cave samples, yet we still obtain low yields. I’ve been developing a “mineral DNA extraction method” based on the principle of repeated freeze-thaw, chemical, and enzymatic lysis with the addition of pyrophosphate to act as a blocking agent and release trapped DNA. To test this new DNA extraction method, Wind cave samples from different locations were supplemented with E.coli and extracted two sets of samples in parallel using the “lab DNA extraction protocol” and the “mineral DNA extraction protocol”. Utilizing the “mineral DNA extraction” obtained a significantly higher yield of DNA compared to our previous lab protocol and enables robust downstream DNA analysis with a more complete sample.
Key words: cave microbiology, DNA extraction, mineral abundant DNA samples Publis
[53a] Hannover Monument #2, Waterloo, Belgium [front]
No full texthttps://scholarworks.uni.edu/his_monuments_sp2022/1102/thumbnail.jp
[62a] Temple Bar, London, England [front]
No full textThis carte-de-visite photograph shows the Temple Bar Gate in London. The gate allowed entrance through the defensive walls surrounding the medieval city. Originally completed in 1672, the gate was taken down in 1878 and reconstructed in a different location. In 2004, the gate was moved back into the city where it remains today.https://scholarworks.uni.edu/his_monuments_sp2022/1120/thumbnail.jp
[63a] Louis XIV Statue, Lyon, France [front]
No full textOriginally put up in 1715, this statue of the “Sun King” was torn down during the French Revolution in 1793. In 1825, it was recast and restored during the Bourbon Restoration, and since then has remained in Lyon’s main square.https://scholarworks.uni.edu/his_monuments_sp2022/1122/thumbnail.jp
[75a] Marcus Aurelius Statue, Rome, Italy [front]
No full textThis carte-de-visite photograph depicts the Equestrian Statue of Marcus Aurelius. The Classical statue was created to honor the Roman Emperor Marcus Aurelius for his triumph in the Marcomannic Wars in c. 175 – 176 C.E. It was moved into the Capitoline Museum in 1981 and replaced by a replica due to corrosion.https://scholarworks.uni.edu/his_monuments_sp2022/1146/thumbnail.jp
[64a] Marshal Louis Suchet Statue, Lyon, France [front]
No full texthttps://scholarworks.uni.edu/his_monuments_sp2022/1124/thumbnail.jp
[76a] Trajan\u27s Column, Rome, Italy [front]
No full textTrajan’s Column in Rome, was constructed to honor Emperor Trajan’s victory in the Dacian Wars. The column was designed and constructed by Apollodorus of Damascus (c. 60-130 C.E.). The monument is 126 feet tall and has a frieze with intricate carvings to depict events during the war.https://scholarworks.uni.edu/his_monuments_sp2022/1148/thumbnail.jp
[78a] The National Monument, Edinburgh, Scotland [front]
No full texthttps://scholarworks.uni.edu/his_monuments_sp2022/1152/thumbnail.jp
Realistic Implementation of OpenSciEd
No full textImmerse yourself into a conversation & real classroom examples on how to make OSE work in your classroom. Come see real examples of how to modify or adjust OSE to best fit SPED accommodations or general population adjustments without losing the rigor